JPS63167313A - Automatic focus control method - Google Patents

Automatic focus control method

Info

Publication number
JPS63167313A
JPS63167313A JP61313824A JP31382486A JPS63167313A JP S63167313 A JPS63167313 A JP S63167313A JP 61313824 A JP61313824 A JP 61313824A JP 31382486 A JP31382486 A JP 31382486A JP S63167313 A JPS63167313 A JP S63167313A
Authority
JP
Japan
Prior art keywords
point
focusing
stage
objective lens
microscope
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
JP61313824A
Other languages
Japanese (ja)
Inventor
Riyouhei Yabe
矢辺　良平
Hajime Matsushita
松下　甫
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hitachi Ltd
Original Assignee
Hitachi Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hitachi Ltd filed Critical Hitachi Ltd
Priority to JP61313824A priority Critical patent/JPS63167313A/en
Priority to US07/136,933 priority patent/US4810869A/en
Publication of JPS63167313A publication Critical patent/JPS63167313A/en
Pending legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/241—Devices for focusing

Abstract

PURPOSE:To rapidly and accurately attain automatic focusing by previously obtaining and storing a focusing position at an optional grid point on a micro scope stage, rapidly moving an objective lens to the obtained approximate focusing position and then executing automatic focusing. CONSTITUTION:A controller 8 executes automatic focusing processing under the state that a practical sample 5 has been previously mounted on an optional grid position on the stage of the microscope 1 and stores the focusing position of the objective lens in each point. Since the focusing positions of the objective lens in respective grid points can be stored once executing the processing, the focusing position of the succeeding sample 5 at a measuring point can be obtained from a grid point near the measuring point. Consequently, the auto matic focusing processing can be executed from a point near the real focusing position of the measuring point.

Description

【発明の詳細な説明】 〔産業上の利用分野〕 本発明は自動焦点制御方法に係り、特に顕微鏡を用いた
パターン認識による細胞自動分類装置に好適な自動焦点
制御方法に関するものである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an automatic focus control method, and more particularly to an automatic focus control method suitable for an automatic cell sorting device using pattern recognition using a microscope.

〔従来の技術〕[Conventional technology]

従来の自動焦点制御は、ある一点における自動焦点を正
確、迅速に行うことを重点において検討されていた。し
かし、高倍率（１００倍）の対物レンズを用いた場合、
その焦点深度は約０．２μＩと非常に浅くなり、顕微鏡
ステージ移動にともなうステージの上下方向のずれに対
する補正については配慮されていなかった。なお、この
種の制御方法に関連するものとして、例えば、特開昭５
９−９４７１１号公報、特開昭５９−１００４０６号公
報等が挙げられる。Conventional automatic focus control has been studied with emphasis on accurately and quickly performing automatic focus at a certain point. However, when using a high magnification (100x) objective lens,
The depth of focus is approximately 0.2 .mu.I, which is very shallow, and no consideration has been given to correction for vertical displacement of the stage as the microscope stage moves. In addition, as related to this type of control method, for example, Japanese Patent Application Laid-open No. 5
9-94711, JP-A-59-100406, and the like.

〔発明が解決しようとする問題点〕[Problem that the invention seeks to solve]

上記従来技術は、顕微鏡ステージを駆動することによる
対物レンズと顕微鏡ステージ間の相対距離の変化による
焦点のボケに対する自動焦点の制御に関して配慮がなさ
れておらず、自動焦点を行うのに時間を要したり、極端
な場合は、自動焦点を実行できないという問題があった
。The above-mentioned conventional technology does not take into consideration automatic focusing control against blurring of focus due to changes in the relative distance between the objective lens and the microscope stage due to driving the microscope stage, and it takes time to perform automatic focusing. In some extreme cases, there was a problem that automatic focusing could not be performed.

本発明の目的は、顕微鏡ステージと対物レンズの相対距
離のずれを補正することにより、迅速、かつ、正確に自
動焦点を行うことができる自動焦点制御方法を提供する
ことにある。An object of the present invention is to provide an automatic focus control method that can quickly and accurately perform automatic focusing by correcting a deviation in the relative distance between a microscope stage and an objective lens.

〔問題点を解決するための手段〕[Means for solving problems]

上記目的は、顕微鏡を用いて観察、測定をする前にあら
かじめ顕微鏡ステージの任意の格子点における合焦点位
置を求めて記憶しておき、実際の測定時には、記憶され
ている近傍の合焦点位置より測定点の合焦点位置を内挿
し、対物レンズを求めた近似合焦点位置に高速移動させ
た後に自動焦点を行うことにより達成するようにした。The above purpose is to determine and memorize the focal point position at an arbitrary grid point on the microscope stage before observing or measuring using a microscope, and then, during actual measurement, from the memorized nearby focal point position. This is achieved by interpolating the in-focus position of the measurement point, moving the objective lens at high speed to the determined approximate in-focus position, and then performing automatic focusing.

〔作用〕[Effect]

制御装置は、あらかじめ顕微鏡ステージの任意の格子点
において実際の標本を架設した状態で自動焦点処理を行
い、各点における対物レンズの合焦点位置を記憶する。The control device performs autofocus processing in advance with an actual specimen set up at an arbitrary grid point on the microscope stage, and stores the in-focus position of the objective lens at each point.

一度この処理を実行すると、各格子点における対物レン
ズの合焦点位置が記憶されるので、以降の標本では、測
定点の近傍の格子点より、８１ＩＩ定点の合焦点位置を
内挿できるので、測定点の真の合焦点位置の近傍より自
動焦点処理を実行することが可能となる。Once this process is executed, the focused position of the objective lens at each grid point is memorized, so for subsequent samples, the focused position of the 81II fixed point can be interpolated from the grid points near the measurement point. It becomes possible to execute automatic focus processing from the vicinity of the true focal point position of a point.

〔実施例〕〔Example〕

以下本発明の制御方法の一実施例を第１図〜第４図を用
いて詳細に説明する。An embodiment of the control method of the present invention will be described in detail below with reference to FIGS. 1 to 4.

第１図は本発明の自動焦点制御方法の一実施例を説明す
るための細胞自動分類装置の一例を示すブロック図であ
る。第１図において、顕微鏡１には光電変換器２および
テレビカメラ３が取り付けられており、顕微鏡１のステ
ージ上に置かれた標本５上に散在する細胞の顕微鏡画像
を光電変換器２、テレビカメラ３に入力する。光電変換
器２から出力された電気信号は、自動焦点装置４に入力
され１合焦点（焦点のピントの一番合ったところ）から
のずれに反比例した焦点信号が出力される。FIG. 1 is a block diagram showing an example of an automatic cell sorting device for explaining an embodiment of the automatic focus control method of the present invention. In FIG. 1, a photoelectric converter 2 and a television camera 3 are attached to a microscope 1, and a microscopic image of cells scattered on a specimen 5 placed on the stage of the microscope 1 is captured by the photoelectric converter 2 and the television camera. Enter 3. The electrical signal output from the photoelectric converter 2 is input to an automatic focusing device 4, which outputs a focus signal that is inversely proportional to the deviation from one in-focus point (the most focused point).

制御袋ｆｉ８はマイクロコンピュータを内蔵しており、
この焦点信号より対物レンズ駆動装置６を制御して、自
動焦点処理を実行する。自動焦点処理が終了すると、テ
レビカメラ３により、細胞の顕微鏡画像の濃度信号が画
像処理装置１７に入力され。The control bag fi8 has a built-in microcomputer,
The objective lens drive device 6 is controlled based on this focus signal to execute automatic focus processing. When the autofocus process is completed, the television camera 3 inputs the density signal of the microscopic image of the cells to the image processing device 17.

画像処理装置！７は制御装置ｌ！８に細胞の特徴パラメ
ータを出力し、制御装置８はこれを基に細胞の分類を行
う６分類が終了すると、ステージ駆動装置９によりステ
ージを移動させ１次の視野内の細胞の分類を続行するか
、または、細胞検出袋Ｗ１１０により対象とする細胞が
検出されるまでステージを移動し１分類を続行する。Image processing device! 7 is the control device l! 8, and the control device 8 classifies the cells based on this. 6 When the classification is completed, the stage is moved by the stage drive device 9 to continue classifying the cells within the primary field of view. Alternatively, the stage is moved and one classification is continued until the target cell is detected by the cell detection bag W110.

分類を実行するにあたり、最初に以下に示す手順でステ
ージ上の各格子点の合焦点位置を記憶する。第２図に示
すように、標本上に任意の間隔ｄ（ｄはステージの精度
に応じて決定し、Ｘ方向。To perform classification, first, the focused position of each grid point on the stage is memorized by the following procedure. As shown in Figure 2, an arbitrary distance d (d is determined according to the precision of the stage) on the specimen in the X direction.

Ｙ方向で必ずしも同じでなくてよい）の格子点をとり、
各格子点における合焦点の対物レンズの位［Ｚ　（ｌ　
ｔ　ｊ）を制御袋Ｗ１８のマイクロコンピュータのメモ
リに記憶する。Take the lattice points (which do not necessarily have to be the same in the Y direction),
The position of the objective lens at the focal point at each grid point [Z (l
t j) is stored in the memory of the microcomputer of the control bag W18.

上記のようにして求められた合焦点位置行列を用いて、
実際に測定を行う場合の手順を以下に述べる。Using the focused position matrix obtained as above,
The procedure for actually performing measurements will be described below.

測定すべき点が第３図に示すＰ点であった場合、その近
傍の格子点で、原点に一番近い格子点からのＸ、Ｙ方向
の距離をΔＸ、Δｙとすると、Ｐ点の位置での合焦点近
似点を次の（１）式で算出する。If the point to be measured is point P shown in Figure 3, let ΔX and Δy be the distances in the X and Y directions from the grid point closest to the origin among the neighboring grid points, then the position of point P is The approximate in-focus point at is calculated using the following equation (1).

Ｚ（Ｐ）＝（（ｄ−Δｘ）・Ｚｚ＋Δｘ−Ｚｚ）／　ｄ
　−（１）ここに、 Ｚｘ＝（（ｄ−Δｙ）・Ｚ（ｘｐｊ）＋Δｙ−Ｚ（ｉ、
ｊ＋１））／ｄＺｚ＝（（ｄ−Δｙ）・ｚ（ｉ＋１．ｊ
）＋Δｙ−Ｚ（ｉ＋１．ｊ＋１））／ｄこのｚ　（ｐ）
の値の位置に対物レンズを移動し、この位置より自動焦
点処理を実行することにより。Z(P)=((d-Δx)・Zz+Δx-Zz)/d
-(1) Here, Zx=((d-Δy)・Z(xpj)+Δy-Z(i,
j+1))/dZz=((d-Δy)・z(i+1.j
)+Δy−Z(i+1.j+1))/dthisz (p)
By moving the objective lens to a value position and performing autofocus processing from this position.

真の合焦点の近傍より自動焦点処理が実行可能となり、
自動焦点処理時間が短縮できる。また、ステージを長い
距離移動した場合、ステージ移動にともない焦点位置が
大幅にずれて自動焦点の実行が不可能になることも防止
可能である。Automatic focus processing can be performed near the true focus point,
Auto focus processing time can be shortened. Furthermore, when the stage is moved over a long distance, it is possible to prevent the focal position from shifting significantly as the stage moves, making it impossible to perform automatic focusing.

なお、第４図は本発明の方法の一実施例を示すフローチ
ャートである。Incidentally, FIG. 4 is a flowchart showing one embodiment of the method of the present invention.

以上述べた実施例では、ステージ移動後に対物レンズを
移動させているが、パルスモータを用いてステージを移
動させるような場合、１パルス移動毎に（１）式を用い
てステージと同時に対物レンズを移動させることも可能
である。In the embodiments described above, the objective lens is moved after the stage is moved, but when the stage is moved using a pulse motor, the objective lens is moved simultaneously with the stage using equation (1) for each pulse movement. It is also possible to move it.

また、合焦点近似点の算出は、（１）式の線形近似だけ
でなく、条件に応じて多項式等の近似を用いることもも
ちろん可能である。Further, in calculating the focal point approximate point, it is of course possible to use not only the linear approximation of equation (1) but also an approximation such as a polynomial depending on the conditions.

本実施例によれば、自動焦点処理を短時間で正確に行う
ことができる。なお、本実施例によるステージは、従来
のものより平滑性においてラフでよく、製作作業が格段
に楽になるという利点がある。According to this embodiment, autofocus processing can be performed accurately in a short time. Note that the stage according to this embodiment has the advantage that its smoothness may be rougher than that of the conventional stage, making the manufacturing work much easier.

〔発明の効果〕〔Effect of the invention〕

以上説明したように、本発明によれば、顕微鏡を用いた
自動焦点制御において、顕微鏡ステージのゆがみを補正
することにより、高速、かつ、正確に自動焦点処理を行
うことができ、高倍率の対物レンズを用いても高精度の
ステージを用いる必要がなく９機構系を安価なものにで
きるという効果がある。As explained above, according to the present invention, in automatic focus control using a microscope, by correcting distortion of the microscope stage, automatic focus processing can be performed quickly and accurately, and high-magnification objective Even if a lens is used, there is no need to use a high-precision stage, and the nine-mechanism system can be made inexpensive.

【図面の簡単な説明】[Brief explanation of the drawing]

第１図は本発明の自動焦点制御方法の一実施例を説明す
るための細胞自動分類装置の一例を示すブロック図、第
２図は補正用格子点の概念図、第３図は補正方法を示す
概念図、第４図は本発明の方法の一実施例を示すフロー
チャートである。 １・・・顕微鏡、２・・・光電変換器、３・・・テレビ
カメラ、４・・・自動焦点装置、５・・・椋本、６・・
・対物レンズ駆動装置、７・・・画像処理装置、８・・
・制御装置、９・・・県１区 ｑ　　−−−ステージ、！３［！ヤ「畏貫帛７ｚ 第３区Fig. 1 is a block diagram showing an example of an automatic cell sorting device for explaining an embodiment of the automatic focus control method of the present invention, Fig. 2 is a conceptual diagram of grid points for correction, and Fig. 3 shows the correction method. The conceptual diagram shown in FIG. 4 is a flowchart showing one embodiment of the method of the present invention. 1... Microscope, 2... Photoelectric converter, 3... Television camera, 4... Automatic focus device, 5... Mukumoto, 6...
・Objective lens drive device, 7... Image processing device, 8...
・Control device, 9...Prefecture 1st ward q---Stage,! 3[! Ya “Awe Penetration 7z Ward 3

Claims (1)

【特許請求の範囲】[Claims] １、顕微鏡の対物レンズを上下方向に駆動する対物レン
ズ駆動装置と、前記顕微鏡のステージを水平方向に駆動
するステージ駆動装置と、前記顕微鏡光量を光電変換し
て得られた電気信号を用いて焦点信号を出力する自動焦
点装置とを備えたものにおいて、前記顕微鏡を用いて観
察、測定をする前にあらかじめ前記顕微鏡ステージの任
意の格子点における合焦点位置を求めて記憶しておき、
実際の測定時には記憶されている近傍の合焦点位置より
測定点の合焦点位置を内挿し、前記対物レンズを求めた
近似合焦点位置に高速移動させた後に自動焦点を行うよ
うにすることを特徴とする自動焦点制御方法。1. An objective lens drive device that drives the objective lens of the microscope in the vertical direction, a stage drive device that drives the stage of the microscope in the horizontal direction, and a focus using an electric signal obtained by photoelectrically converting the amount of light from the microscope. and an automatic focusing device that outputs a signal, in which a focused point position at an arbitrary grid point of the microscope stage is determined and stored in advance before observation and measurement using the microscope;
During actual measurement, the in-focus position of the measurement point is interpolated from the memorized nearby in-focus position, and the objective lens is moved at high speed to the determined approximate in-focus position, and then automatic focusing is performed. automatic focus control method.
JP61313824A 1986-12-27 1986-12-27 Automatic focus control method Pending JPS63167313A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP61313824A JPS63167313A (en) 1986-12-27 1986-12-27 Automatic focus control method
US07/136,933 US4810869A (en) 1986-12-27 1987-12-23 Automatic focusing control method for microscope

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP61313824A JPS63167313A (en) 1986-12-27 1986-12-27 Automatic focus control method

Publications (1)

Publication Number Publication Date
JPS63167313A true JPS63167313A (en) 1988-07-11

Family

ID=18045953

Family Applications (1)

Application Number Title Priority Date Filing Date
JP61313824A Pending JPS63167313A (en) 1986-12-27 1986-12-27 Automatic focus control method

Country Status (2)

Country Link
US (1) US4810869A (en)
JP (1) JPS63167313A (en)

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